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. 2012 May 9;32(19):6468–6484. doi: 10.1523/JNEUROSCI.0590-12.2012

Figure 1.

Figure 1.

gfp knock-in mice for the Apc2 gene. A, Schematic representation of the structure of the endogenous allele (genomic), targeting vector (vector), and targeted allele (targeted). The genomic sequence of the head region of exon 2 in the targeted allele is shown below together with the encoded amino acid sequence. The protein coding exons of Apc2 are indicated by black boxes. For construction of the targeting vector, an egfp-Neo cassette was inserted in frame in the signal sequence of APC2 after the first two amino acids, Met-Ala, to yield fusion to the N terminus of GFP through a linker sequence. The diphtheria toxin A (DTA) gene cassette was placed at the 3′ terminus of the homologous region in the targeting vector for negative selection. The region used as a probe for Southern blotting is indicated by a bold bar. B, Southern blot analysis of SalI and BamHI double-digested genomic DNA from wild-type (+/+), heterozygous (+/−), and homozygous (−/−) mice. C, RT-PCR analysis of total RNA from the brain of wild-type (+/+), heterozygous (+/−), and homozygous (−/−) mice. D, Western blot analysis of brain extracts from wild-type (+/+), heterozygous (+/−), and homozygous (−/−) mice using an anti-APC2, GFP, or GAPDH antibody. E, Wild-type and homozygous Apc2-deficient E14.5 embryos. F, Wild-type and homozygous Apc2-deficient P180 mice. Scale bars: E, 1 mm; F, 2 mm.